title: Automated procedure for biomimetic de-cellularized lung scaffold supporting alveolar epithelial transdifferentiation.
abstract: The optimal method for creating a de-cellularized lung scaffold that is devoid of cells and cell debris, immunologically inert, and retains necessary extracellular matrix (ECM) has yet to be identified. Herein, we compare automated detergent-based de-cellularization approaches utilizing either constant pressure (CP) or constant flow (CF), to previously published protocols utilizing manual pressure (MP) to instill and rinse out the de-cellularization agents. De-cellularized lungs resulting from each method were evaluated for presence of remaining ECM proteins and immunostimulatory material such as nucleic acids and intracellular material. Our results demonstrate that the CP and MP approaches more effectively remove cellular materials but differentially retain ECM proteins. The CP method has the added benefit of being a faster, reproducible de-cellularization process. To assess the functional ability of the de-cellularized scaffolds to maintain epithelial cells, intra-tracheal inoculation with GFP expressing C10 alveolar epithelial cells (AEC) was performed. Notably, the CP de-cellularized lungs were able to support growth and spontaneous differentiation of C10-GFP cells from a type II-like phenotype to a type I-like phenotype.
authors.affiliation: Department of Surgery, Connecticut Children's Medical Center, 282 Washington Street, Hartford, CT 06106, USA; Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue, MC3501, Farmington, CT 06030, USA.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Girard
authors.forename: Todd J
authors.initials: TJ
authors.lastname: Jensen
authors.forename: Stephanie D
authors.initials: SD
authors.lastname: Vadasz
authors.forename: Alex E
authors.initials: AE
authors.lastname: Blanchette
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.forename: Camilo
authors.initials: C
authors.lastname: Moncada
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Weiss
authors.forename: Christine M
authors.initials: CM
authors.lastname: Finck
mesh_terms: D000818:Animals; D032701:Biomimetics; D054338:Cell Transdifferentiation / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D008168:Lung / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D054457:Tissue Scaffolds / Q000737:chemistry
keywords: Alveolar epithelial cell; C10 alveolar cells; De-cellularization; Extracellular matrix; Lung; Scaffold